CN107402209B - Test strip for detecting low-density lipoprotein cholesterol in serum and preparation method thereof - Google Patents
Test strip for detecting low-density lipoprotein cholesterol in serum and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a test strip for detecting low-density lipoprotein cholesterol in serum, which comprises a dry chemical reaction sheet, wherein the dry chemical reaction sheet is prepared by soaking a reagent pad in a chemical reagent and drying, and the chemical reagent adopts cholesterol esterase, cholesterol oxidase, peroxidase, metal salt, a color developing agent, polyoxyethylene ether, ethylene oxide-propylene oxide copolymer and the like with optimal concentration and proportion. The method has the advantages of low equipment cost, easy preparation and acquisition, quick reaction and convenient operation.
Description
Technical Field
The invention relates to a test strip for detecting low-density lipoprotein cholesterol in serum and a preparation method thereof.
Background
Abnormal lipid metabolism is one of the most important risk factors for Coronary Heart Disease (CHD). Numerous large-scale clinical studies have well documented the important role of cholesterol, particularly low-density lipoprotein cholesterol (LDL-C), in the development of coronary heart disease, and lowering LDL-C can significantly reduce coronary heart disease events. The rise of low density lipoprotein cholesterol is in positive correlation with the incidence of coronary heart disease, and is one of the important analysis indexes of coronary heart disease. Traditional LDL-C concentrations are obtained by means of separate assays or indirect approximate calculations. The separation and measurement are carried out by separating lipoprotein components in blood samples according to high density lipoprotein, low density lipoprotein and chylomicron by ultracentrifuge according to certain steps, and then measuring separately. The method has high requirement on equipment, long time consumption and complex operation. While the indirect approximation calculation mode is to calculate the approximate value of LDL-C by a Friedewald formula according to the values of Total Cholesterol (TC), Triglyceride (TG) and high density lipoprotein cholesterol (HDL-C), the method is only suitable for TG <4.52 mmol/L. When CM and TG are high in serum, calculated values are large in deviation, and a plurality of factors influence the accuracy of the formula. Therefore, only direct measurement of LDL-C can provide accurate diagnosis basis for clinical application. Recent years have seen the direct detection of LDL-c on fully automated biochemists. The scheme is realized mainly by protecting HDL from reaction or delaying the reaction speed of HDL part in an enzyme system of a traditional total cholesterol detection reaction through some special surfactants, so as to achieve the aim that only LDL participates in the enzyme system reaction, and finally only the concentration detection result of cholesterol in LDL, namely the concentration of LDL-c in a sample is obtained.
So far, the direct method for detecting LDL-c in serum samples mainly comprises two methods, namely a selective inhibition method and an elimination method, and the specific technical schemes of the two methods are as follows:
1. selective inhibition method: the method is a method of measuring the amount of LDL-C alone hydrolyzed by a cholesterol esterase and a cholesterol oxidase by inhibiting the reaction (also referred to as selective inhibition) of non-LDL-C lipoproteins such as HDL and VLDL in a serum sample with the use of a-cyclodextrin, dextran sulfate, Brij58, and the like.
A two-reagent reaction system is typically used with this method. The reagent 1 contains alpha-cyclodextrin sodium sulfate, dextran sulfate and MgCl2EMSE and PBS. Reagent 2 contains CHER, CHOD peroxidase, 4-AA, surfactant and PBS. Sodium a-cyclodextrin sulfate in small amount of DS and Mg2+In the presence of a specific surfactant, the reaction of cholesterol in the Chylomicron (CM) and VLDL components with the enzymatic reagent is reduced, and the reaction of cholesterol in the HDLC component is reduced, and in combination, LDL-c can be selectively measured.
2. Elimination method: the two-step hydrolysis of non-LDL-c and LDL-c with two reagents of different surfactants, which is called elimination method because non-LDL-c is consumed first, is a direct measurement method which is widely applied at present.
The method also employs a two-reagent reaction system. Mainly comprises a surfactant elimination method, a catalase elimination method and the like. The reagent of the first chemical Genxyme Diagnostics company of Japan belongs to the method, and is the most widely used reagent at home and abroad. The surfactant 1 in the reagent 1 can change the structures of HDL, CM and VLDL except LDL-c and dissociate, and the released micronized cholesterol molecules react with the cholesterol enzyme reagent to generate H2O2Is consumed without developing color in the absence of coupling agent, and the LDL-c particles are dissociated to release cholesterolIt takes part in enzyme reaction to develop color, and the color depth is proportional to the amount of LDL-c.
The two detection methods both belong to two-component biochemical reagents, and both require a spectrophotometer or a full-automatic biochemical analyzer to execute relevant reactions and perform result calculation analysis.
Therefore, the above-mentioned techniques have problems in that the measurement of LDL-c concentration involves: the scheme of separating LDL and HDL by ultracentrifugation needs to use an expensive ultra-high speed centrifuge as a main separation device, and has the disadvantages of multiple experimental steps, complicated operation, preparation of various reagents, most importantly, time consumption, and the whole separation time often exceeds 6 hours. The direct detection method requires large-scale automation equipment such as a full-automatic biochemical analyzer, and is high in cost.
A method for measuring low-density lipoprotein cholesterol having an authorization publication No. CN101864475B, an authorization publication date of 2013-11-6, which discloses a dry analytical element for measuring LDL-c, which is prepared by coating a gelatin aqueous solution on a colorless, transparent, smooth film of polyethylene terephthalate having a thickness of 180 micrometers, which is coated with gelatin on the lower side, and drying until the dried thickness reaches 14 micrometers, coating an aqueous solution having the following composition, and then drying. 4-aminoantipyrine 0.32g/m2、TOOS 0.62 g/m2Peroxidase 12.75 KU/m2. About 30 m above the film2The amount of (c) was such that each side was moistened with water. Then, a light-knitted fabric obtained by knitting a polyester spun yarn equivalent to 50 denier in a 36-gauge was laminated with a light pressure and dried. Next, an aqueous solution having the following composition was applied to the cloth, followed by drying. MOPS (pH7.0) 1.67 g/m25.02 g/m polyglycerol tristyryl phenyl ether2、Pluronic L 121 39.75 g/m20.65 KU/m cholesterol esterase (lipoprotein lipase, Toyobo)2Cholesterol oxidase (recombinant Escherichia coli, Kikkoman) 0.13 KU/m2. It can be seen that the dry analytical element made in this patent requires two reagents to be fixed to different films, respectively, and thus, the manufacturing process is cumbersome.
Disclosure of Invention
The technical problem to be solved by the invention is to provide the test strip for detecting low-density lipoprotein cholesterol in serum, which has low cost and simple preparation.
The invention also provides a preparation method of the test strip.
In order to solve the technical problems, the invention adopts the following technical scheme:
the invention aims to provide a test strip for detecting low-density lipoprotein cholesterol in serum, which comprises a dry chemical reaction sheet, wherein the dry chemical reaction sheet is prepared by soaking a reagent pad in a chemical reagent and then drying, and the chemical reagent comprises cholesterol esterase with the concentration of 1-30 KU/L, cholesterol oxidase with the concentration of 20-80 KU/L, 250-350 mM metal salt, 100-400 KU/L horseradish peroxidase, 15-25 mM 4-aminoantipyrine, 40-60 mM color developing agent, 0.12-0.2 g/mL polyoxyethylene ether surfactant, 1.5-2.5 g/L ethylene oxide-propylene oxide copolymer surfactant and phosphate buffer.
Preferably, the chemical reagent comprises cholesterol esterase with the concentration of 5-15 KU/L, 40-60 KU/L cholesterol oxidase, 280-320 mM metal salt, 150-250 KU/L horseradish peroxidase, 18-22 mM 4-aminoantipyrine, 45-55 mM color developing agent, 0.14-0.18 g/mL polyoxyethylene ether surfactant, 1.8-2.2 g/L ethylene oxide-propylene oxide copolymer surfactant and phosphate buffer.
More preferably, the chemical reagent comprises cholesterol esterase with the concentration of 5-8 KU/L, 40-45 KU/L cholesterol oxidase, 280-290 mM metal salt, 150-180 KU/L horseradish peroxidase, 18-19 mM 4-aminoantipyrine, 45-48 mM color developing agent, 0.14-0.15 g/mL polyoxyethylene ether surfactant, 1.8-1.9 g/L ethylene oxide-propylene oxide copolymer surfactant and phosphate buffer.
Preferably, the metal salt is one or more selected from magnesium sulfate, magnesium chloride, sodium sulfate, sodium chloride, calcium chloride, magnesium phosphate and sodium phosphate; further preferred is magnesium sulfate or magnesium chloride.
Preferably, the developer is a developer selected from the group consisting of N- (2-hydroxy-3-sulfopropyl) -35-dimethoxyaniline sodium salt (HDAOS), N-ethyl-N- (2-hydroxy-3-propanesulfo) m-Toluidine (TOOS), N-bis (4-sulfopropyl) -3, 5-dimethylaniline sodium salt (MADB), N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-dimethylaniline sodium salt Monohydrate (MAOS), N-ethyl-N- (3-sulfopropyl) -3-methoxyaniline sodium salt (ADPS), N-bis (4-sulfobutyl) -3-methylaniline disodium salt (TODB), One or more of N-ethyl-N- (3-sulfopropyl) -3-methylaniline sodium salt (TOPS), N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-dimethoxyaniline sodium salt (DAOS), nitro blue tetrazolium chloride (NBT), iodine nitro blue tetrazolium chloride (INT) and blue tetrazolium chloride (BT); further preferred is N-ethyl-N- (2-hydroxy-3-propanesulfo) m-Toluidine (TOOS).
Preferably, the polyoxyethylene ether surfactant is one or more selected from fatty alcohol polyoxyethylene ether, nonylphenol polyoxyethylene ether, sorbitan monooleate polyoxyethylene ether and polyoxyethylene lauryl ether.
Further preferably, the fatty alcohol-polyoxyethylene ether is one or more selected from AEO-3, AEO-5, AEO-7 and AEO-10.
Further preferably, the polyoxyethylene nonyl phenyl ether is one or more selected from NP-4, NP-10 and NP-15.
Further preferably, the polyoxyethylene sorbitan monooleate is tween-20 and/or tween-80.
Further preferably, the polyoxyethylene lauryl ether is Brij-35 and/or Brij-58.
Preferably, the number average molecular weight of the ethylene oxide-propylene oxide copolymer surfactant is 5000-15000.
Further preferably, the number average molecular weight of the ethylene oxide-propylene oxide copolymer surfactant is 6000-10000.
More preferably, the number average molecular weight of the ethylene oxide-propylene oxide copolymer surfactant is 8000-9000.
Preferably, the ethylene oxide-propylene oxide copolymer surfactant is one or more selected from poloxamer F124, poloxamer F88, Pluronic L-122 and Pluronic L-101.
Preferably, the material of the reagent pad is one of a nitrocellulose membrane, a glass cellulose membrane, a polyvinylidene fluoride cellulose membrane, a polyacetate cellulose membrane, a polysulfonamide membrane and a polyethyleneimine membrane. More preferably, a cellulose acetate film.
Preferably, the pore size of the reagent pad is 0.3 to 0.5 micron, more preferably 0.35 to 0.45 micron, and most preferably 0.4 micron. Preferably, the test strip further comprises a sample release pad lining the dry chemistry reaction chip.
Further preferably, the material of the sample release pad is a glass fiber film.
Further preferably, the thickness of the sample release pad is 1 to 3 mm, more preferably 1.5 to 2.5 mm, and most preferably 2 mm.
The invention also aims to provide a preparation method of the test strip for detecting low-density lipoprotein cholesterol in serum, which comprises the following steps:
mixing the cholesterol esterase, the cholesterol oxidase, the metal salt, the horseradish peroxidase, the 4-aminoantipyrine, the color developing agent and the phosphate buffer solution to form a mixed solution;
adding the polyoxyethylene ether surfactant and the ethylene oxide-propylene oxide copolymer surfactant into the mixed solution, and mixing to form the chemical reagent;
and (3) placing the reagent pad into the chemical reagent, soaking for 60-90 min, and then drying for 40-60 min at 45-50 ℃ to obtain the dry chemical reaction plate.
Preferably, the preparation method further comprises the step of lining the sample release liner on the dry chemistry reaction chip.
Due to the implementation of the technical scheme, compared with the prior art, the invention has the following advantages:
the invention provides a test strip for detecting low-density lipoprotein cholesterol in serum based on a dry chemical method, which is prepared by soaking a reagent pad in a chemical reagent and then drying the reagent pad without using two reagents to be loaded on different films respectively.
Drawings
FIG. 1 is a graph showing the correlation between the results of the LDL-c dry chemical reaction plate of example 1 and the results of the full-automatic biochemical analyzer;
FIG. 2 is a graph showing the correlation between the results of the LDL-c dry chemical reaction plate of example 2 and the results of the full-automatic biochemical analyzer;
FIG. 3 is a graph showing the correlation between the results of the LDL-c dry chemical reaction plate of example 3 and the results of the full-automatic biochemical analyzer.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the present invention is not limited to the following examples. The implementation conditions adopted in the embodiments can be further adjusted according to different requirements of specific use, and the implementation conditions not mentioned are conventional conditions in the industry. All other embodiments obtained by a person skilled in the art without making any inventive step are within the scope of protection of the present invention.
The reagents used in the present invention are commercially available.
Example 1:
(1) preparing a Total Cholesterol (TC) detection reagent: according to cholesterol esterase (CHER) 5KU/L, cholesterol oxidase (CHOD) 40KU/L, MgSO4280mM, 150KU/L of horseradish peroxidase, 18 mM of 4-aminoantipyrine, and 45 mM of N-ethyl-N- (2-hydroxy-3-propanesulfo) m-Toluidine (TOOS), and phosphoric acid (PBS) as a buffer, all of the above reagents were added to a 15mL EP tube and mixed well.
) 0.14g of Tween-20 (Tween-20) is added into the reagent obtained in the first step according to 1mL of the reagent, and then poloxamer F124 with the number average molecular weight of 8400 is added according to the concentration of 1.8g/L and is uniformly mixed.
) Cutting the cellulose nitrate film with the aperture of 0.4um into a strip with the width of about 0.5cm, putting the strip into the solution prepared in the previous step, and soaking for 1.5 hours. Then taking out, putting into an electric heating air blast drying oven, and drying for 60min at the temperature of 45 ℃. The completely dried membrane strip is the LDL-c dry chemical reaction sheet.
) A test strip was prepared using a 2mm thick glass fiber membrane as a sample release pad, lined on an LDL-c dry chemistry reaction plate.
(5) And dropwise adding the serum to be detected on the sample release pad by using a pipette, uniformly soaking the LDL-c dry chemical reaction sheet by the serum through the sample release pad, and changing color along with the reaction to realize the dry chemical detection of the LDL-c.
(6) The test strip of this example was used to perform LDL-c assay on clinical serum samples using the reagent of first chemistry Genxyme Diagnostics, Japan, respectively, using a fully automatic biochemical analyzer, and the results are shown in FIG. 1, where FIG. 1 is the correlation data between the assay results of the LDL-c dry chemistry reaction plate and the assay results of the fully automatic biochemical analyzer, and it can be seen that the assay results of the LDL-c dry chemistry reaction plate and the assay results of the fully automatic biochemical analyzer have a good correlation.
Example 2:
(1) preparing a Total Cholesterol (TC) detection reagent: according to 15KU/L of cholesterol esterase (CHER), 60KU/L of cholesterol oxidase (CHOD), MgSO4320mM, 250KU/L of horseradish peroxidase, 22mM of 4-aminoantipyrine, and 55mM of N-ethyl-N- (3-sulfopropyl) -3-methylaniline sodium salt (TOPS) were added to a 15mL EP tube using phosphoric acid (PBS) as a buffer solution, and the above reagents were mixed well.
) 0.18g of Tween-20 (Tween-20) is added into the reagent obtained in the first step according to 1mL of the reagent, and then poloxamer F88 with the number average molecular weight of 8400 is added according to the concentration of 2.2g/L and is uniformly mixed.
) Cutting the cellulose nitrate film with the aperture of 0.4um into a strip with the width of about 0.5cm, putting the strip into the solution prepared in the previous step, and soaking for 1.5 hours. Then taking out, putting into an electric heating air blast drying oven, and drying for 60min at the temperature of 45 ℃. The completely dried membrane strip is the LDL-c dry chemical reaction sheet.
) A test strip was prepared using a 2mm thick glass fiber membrane as a sample release pad, lined on an LDL-c dry chemistry reaction plate.
(5) And dropwise adding the serum to be detected on the sample release pad by using a pipette, uniformly soaking the LDL-c dry chemical reaction sheet by the serum through the sample release pad, and changing color along with the reaction to realize the dry chemical detection of the LDL-c.
(6) The test strip of this example and the reagent of first chemical Genxyme Diagnostics of Japan were used to perform LDL-c detection on five clinical serum samples with a fully automatic biochemical analyzer, and the results are shown in FIG. 2. FIG. 2 is the correlation data between the detection results of the LDL-c dry chemical reaction plate and the detection results of the fully automatic biochemical analyzer, and it can be seen that the detection results of the LDL-c dry chemical reaction plate and the detection results of the fully automatic biochemical analyzer have a good correlation.
Example 3:
(1) preparing a Total Cholesterol (TC) detection reagent: according to 10KU/L of cholesterol lipase (CHER), 50KU/L of cholesterol oxidase (CHOD), MgCl2300mM, 200KU/L horseradish peroxidase, 20mM 4-aminoantipyrine and 50mM nitrotetrazolium chloride (NBT) are added into a 15mL EP tube by taking phosphoric acid (PBS) as a buffer solution and mixed uniformly.
) Adding 0.16g NP-10 into the reagent obtained in the first step according to 1mL of the reagent, adding poloxamer F88 with the number average molecular weight of 6000 into the reagent according to the concentration of 2g/L, and uniformly mixing.
) Cutting the cellulose nitrate film with the aperture of 0.4um into a strip with the width of about 0.5cm, putting the strip into the solution prepared in the previous step, and soaking for 1.5 hours. Then taking out, putting into an electric heating air blast drying oven, and drying for 60min at the temperature of 45 ℃. The completely dried membrane strip is the LDL-c dry chemical reaction sheet.
) A test strip was prepared using a 2mm thick glass fiber membrane as a sample release pad, lined on an LDL-c dry chemistry reaction plate.
(5) And dropwise adding the serum to be detected on the sample release pad by using a pipette, uniformly soaking the LDL-c dry chemical reaction sheet by the serum through the sample release pad, and changing color along with the reaction to realize the dry chemical detection of the LDL-c.
(6) The test strip of this example was used to test LDL-c in clinical serum samples using the reagent of Genxyme Diagnostics, Japan, using a fully automatic biochemical analyzer, and the results are shown in FIG. 3, from which it can be seen that the results of the test of the LDL-c dry chemistry reaction plate are better correlated with the results of the fully automatic biochemical analyzer.
The present invention has been described in detail in order to enable those skilled in the art to understand the invention and to practice it, and it is not intended to limit the scope of the invention, and all equivalent changes and modifications made according to the spirit of the present invention should be covered by the present invention.
Claims (9)
1. A test strip for detecting low density lipoprotein cholesterol in serum is characterized in that: the test strip comprises a dry chemical reaction sheet and a sample release pad lined on the dry chemical reaction sheet, wherein the dry chemical reaction sheet is prepared by soaking a reagent pad in a chemical reagent and then drying, and the chemical reagent comprises cholesterol esterase with the concentration of 1-30 KU/L, 20-80 KU/L cholesterol oxidase, 250-350 mM metal salt, 100-400 KU/L horseradish peroxidase, 15-25 mM 4-aminoantipyrine, 40-60 mM color developing agent, 0.12-0.2 g/mL polyoxyethylene ether surfactant, 1.5-2.5 g/L ethylene oxide-propylene oxide copolymer surfactant and a phosphate buffer solution; the metal salt is one or more selected from magnesium sulfate, magnesium chloride, sodium sulfate, sodium chloride, calcium chloride, magnesium phosphate and sodium phosphate; the polyoxyethylene ether surfactant is one or more selected from fatty alcohol polyoxyethylene ether, nonylphenol polyoxyethylene ether, sorbitan monooleate polyoxyethylene ether and polyoxyethylene lauryl ether.
2. The test strip for detecting low density lipoprotein cholesterol in serum according to claim 1, wherein: the chemical reagent comprises cholesterol esterase with the concentration of 5-15 KU/L, 40-60 KU/L cholesterol oxidase, 280-320 mM metal salt, 150-250 KU/L horseradish peroxidase, 18-22 mM 4-aminoantipyrine, 45-55 mM color developing agent, 0.14-0.18 g/mL polyoxyethylene ether surfactant, 1.8-2.2 g/L ethylene oxide-propylene oxide copolymer surfactant and phosphate buffer.
3. The test strip for detecting low density lipoprotein cholesterol in serum according to claim 1 or 2, characterized in that:
the color developing agent is selected from N- (2-hydroxy-3-sulfopropyl) -35-dimethoxyaniline sodium salt, N-ethyl-N- (2-hydroxy-3-propanesulfo) m-toluidine, N-di (4-sulfopropyl) -3, 5-dimethylaniline sodium salt, N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-dimethylaniline sodium salt monohydrate, N-ethyl-N- (3-sulfopropyl) -3-methoxyaniline sodium salt, N-di (4-sulfobutyl) -3-methylaniline disodium salt, N-ethyl-N- (3-sulfopropyl) -3-methylaniline sodium salt, One or more of N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3, 5-dimethoxyaniline sodium salt, nitro blue tetrazolium chloride iodide and blue tetrazolium chloride.
4. The test strip for detecting low density lipoprotein cholesterol in serum according to claim 1 or 2, characterized in that: the number average molecular weight of the ethylene oxide-propylene oxide copolymer surfactant is 5000-15000.
5. The test strip for detecting low density lipoprotein cholesterol in serum according to claim 4, wherein: the number average molecular weight of the ethylene oxide-propylene oxide copolymer surfactant is 6000-10000.
6. The test strip for detecting low density lipoprotein cholesterol in serum according to claim 1 or 2, characterized in that: the material of the reagent pad is one of a nitrocellulose membrane, a glass cellulose membrane, a polyvinylidene fluoride cellulose membrane, a polyacetate cellulose membrane, a polysulfonamide membrane and a polyethyleneimine membrane.
7. The test strip for detecting low density lipoprotein cholesterol in serum according to claim 1, wherein: the sample release pad is made of a glass fiber film, and the thickness of the glass fiber film is 1-3 mm.
8. A method for preparing the test strip for detecting low density lipoprotein cholesterol in serum according to any one of claims 1 to 7, which comprises: the method comprises the following steps:
mixing the cholesterol esterase, the cholesterol oxidase, the metal salt, the horseradish peroxidase, the color-developing agent, the 4-aminoantipyrine and the phosphate buffer solution to form a mixed solution;
adding the polyoxyethylene ether surfactant and the ethylene oxide-propylene oxide copolymer surfactant into the mixed solution, and mixing to form the chemical reagent;
and (3) placing the reagent pad into the chemical reagent, soaking for 60-90 min, and then drying for 40-60 min at 45-50 ℃ to obtain the dry chemical reaction plate.
9. The method of claim 8, wherein: the preparation method further comprises the step of lining the sample release liner on the dry chemistry reaction chip.
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| CN110029145A (en) * | 2018-01-11 | 2019-07-19 | 北京瑞正善达生物工程技术有限公司 | Low density lipoprotein cholesterol measures reagent, preparation method and its application method |
| CN108535243B (en) * | 2018-03-26 | 2020-12-01 | 北京普赞生物技术有限公司 | Color development test paper for rapid detection of cyanide and preparation and detection methods thereof |
| CN109097436A (en) * | 2018-08-10 | 2018-12-28 | 山东博科生物产业有限公司 | A kind of low density lipoprotein cholesterol detection reagent of the single agents of precise and high efficiency |
| CN109916890A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining total cholesterol concentration |
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| CN102539731A (en) * | 2012-01-09 | 2012-07-04 | 宁波天康生物科技有限公司 | Reagent and kit for quantitatively determining low-density lipoprotein cholesterol (LDL-C) in serum |
| CN104076158A (en) * | 2014-07-10 | 2014-10-01 | 深圳市新产业生物医学工程股份有限公司 | Kit for measuring high-density lipoprotein cholesterol |
| CN106199018A (en) * | 2016-07-07 | 2016-12-07 | 长春理工大学 | A kind of monofilm biochemistry dry tablet for detecting blood fat and preparation method |
| CN106645763A (en) * | 2017-01-03 | 2017-05-10 | 长沙中生众捷生物技术有限公司 | Total cholesterol detection reagent and total cholesterol detection paper |
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